Ezrin ubiquitylation by WWP1 is required for Met upregulation.

<p>(A) Met levels in LLC-PK1 cells expressing GFP-WWP1 or GFP alone. Left panel: Blots were performed with anti-Met and anti-tubulin antibodies. Right panel: The quantification of Met level normalized to tubulin results from five independent experiments. Data are expressed as mean ± SEM, *P<0.05 (paired t-test). (B) Ezrin depletion prevents Met upregulation. Stable GFP-WWP1 cells were transfected with plasmids coding for shRNA targeting ezrin (shEz) or a scramble sequence (Scr). Left panel: Cell lysates were blotted with the indicated antibodies. Right panel: the quantification of Met level normalized to tubulin results from five independent experiments. Data are expressed as mean ± SEM, *P<0.05 (paired t-test). (C) Cell speed measured in a wound healing assay. In each graph, the bars represent the increase in the speed of cells treated with HGF as compared to the speed of untreated cells. Left panel: LLC-PK1 cells expressing either GFP or GFP-WWP1. Middle panel: LLC-PK1 cells expressing GFP-WWP1 and transfected with plasmids coding for scramble shRNA (Scr) or shRNA targeting ezrin. Right panel: LLC-PK1 cells expressing GFP-WWP1 and shRNA targeting ezrin were transfected with either an empty vector (vector) or plasmids coding for ezrin wild type or ezrin P475A. The data correspond to three independent experiments. Significance was tested using a two-way ANOVA model with interaction (SigmaStat). The pairwise comparisons were performed with the Student-Newman-Keuls method using an α risk of 0.05.